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Abstract:

The present invention provides a method for treating inflammation in a
patient in need thereof comprising administering to said patient an
effective amount of a compound according to formula
##STR00001##
wherein R is H or lower alkyl, R1 is hydrocarbyl or substituted
hydrocarbyl and the broken line represents a saturated or unsaturated
bond, i.e a double bond. Preferably, R1 is an alkyl. More preferably, R1
is a n-alkyl or a cycloalkyl-n-alkyl, e.g. a cyclohexyl-n-alkyl, e.g.
n-octyl, n-nonyl or cyclohexyl-n-butyl radical and prodrugs, isomers and
pharmaceutically acceptable salts thereof.

Claims:

1. A method for treating inflammation in a patient in need thereof
comprising administering to said patient an effective amount of a
compound according to formula ##STR00039## wherein R is H or lower
alkyl, R1 is hydrocarbyl or substituted hydrocarbyl and the broken line
represents a saturated or unsaturated bond, or a double bond, wherein
said formula further including prodrugs, isomers and pharmaceutically
acceptable salts thereof.

2. The method of claim 1 wherein said compound is ##STR00040##

3. The method of claim 2 wherein said method comprises decreasing the
secretion of ENA-78 in a patient.

4. The method of claim 2 wherein said method comprises decreasing the
secretion of IL-8 in a patient.

5. The method of claim 2 wherein said method comprises decreasing the
secretion of MCP-1 in a patient.

6. The method of claim 2 wherein said method comprises decreasing the
secretion of PAI-1 in a patient.

7. The method of claim 2 wherein said method comprises decreasing the
secretion of CD-40 in a patient.

8. The method of claim 2 wherein said method comprises decreasing the
secretion of G-C--SF in a patient.

9. The method of claim 2 wherein said method comprises decreasing the
secretion of GM-CSF in a patient.

10. The method of claim 2 wherein said method comprises decreasing the
secretion of IL-1.alpha. in a patient.

11. The method of claim 2 wherein said method comprises decreasing the
secretion of IL-18 in a patient.

12. The method of claim 2 wherein said method comprises decreasing the
secretion of MDC in a patient.

13. The method of claim 2 wherein said method comprises decreasing the
secretion of RANTES in a patient.

14. The method of claim 2 wherein said compound is at least as effective
as COXIBs and NSAIDs in treating inflammation in a patient in need of
such treatment, without causing cardiovascular, renal and/or
gastro-intestinal side effects.

15. A method for decreasing the secretion of the cytokine selected from
the group consisting of ENA-78, IL-8, MCP-1, PAI-1, TNFα, CD-40,
G-CSF, GM-CSF, IL-1.alpha., IL-18, MDC and RANTES in a patient in need
thereof comprising administering to a patient an effective amount of a
compound according to formula I: ##STR00041## wherein R is H or lower
alkyl, R1 is hydrocarbyl or substituted hydrocarbyl and the broken line
represents a saturated or unsaturated bond, and a double bond.

16. The method of claim 15 wherein said compound is ##STR00042##

17. A method according to claim 15 wherein said cytokine is ENA-78 and
said compound is administered for treating rheumatoid arthritis.

18. A method according to claim 15 wherein said cytokine is IL-8 and said
compound is administered for treating rheumatoid arthritis.

19. A method according to claim 15 wherein said cytokine is MCP-1 and
said compound is administered for treating inflammatory diseases
characterized by monocytic infiltration, such as diseases selected from
the group consisting of RA rheumatoid arthritis, psoriasis, and
atherosclerosis; atopic dermatitis, renal disease; pleurisy; allergy and
asthma; colitis; endometriosis; polymyositis and dermatomyositis;
uveitis; restenosis; brain inflammation and obesity; diabetes and
diabetes-induced atherosclerosis and MCP-1/CCR2-mediated multiple
inflammatory diseases.

20. A method according to claim 15 wherein said cytokine is CD-40 and
said compound is administered for treating rheumatoid arthritis,
inflammation, thrombosis and atherosclerosis, chronic renal failure,
chronic liver diseases, Alzheimer's disease and systemic sclerosis.

Description:

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent
Application Ser. Nos. 61/360,738 filed on Jul. 1, 2010 and U.S.
Provisional Patent Application Ser. Nos. 61/410,153 filed on Nov. 4, 2010
both of which are hereby incorporated by reference in its entirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to compounds that show an
anti-inflammatory action and are useful as pharmaceuticals

[0004] 2. Summary of the Related Art

[0005] At present, the majority of medicines widely used as
anti-inflammatory agents are non-steroid anti-inflammatory drugs (NSAIDs)
that have, as the mechanism of action, an inhibitory action on
cyclooxygenases (COXs) that is involved in the biosynthesis of
prostanoids. However, since prostanoid synthesis activity is present in
various tissues in the living body and governs the homeostasis thereof,
various side effects are induced when NSAID is administered. For example,
PGE2 demonstrates the action of maintaining blood flow in the stomach and
kidneys, whereas administration of NSAIDs makes it difficult to maintain
local blood flow, thereby causing gastric or renal disorders.

[0006] Under such circumstances, the presence of a COX isozyme has been
confirmed. In order to distinguish it from the previously identified COX,
the conventional type has been named COX-1, while the more recently
discovered isozyme has been named COX-2. In addition, this COX-2 has been
shown to be induced during inflammation and hardly be expressed under
normal circumstances. It has also been shown that conventional NSAID are
able to non-specifically inhibit both COX-1 and COX-2 enzymes. Therefore,
a compound having COX-2 inhibitory action would be useful as an
anti-inflammatory agent.

[0007] There are currently several compounds that are known to
preferentially inhibit COX-2 whilst having significantly less COX-1
inhibitory activity. However, the actions of these compounds are not
satisfactory and since some of them do not have an adequate water
solubility or oral absorption, there remains a need for a drug that
demonstrates more effective COX-2 inhibitory action.

[0008] Vioxx or 4-[4-(methylsulfonyl)phenyl]-3-phenyl-2(5H)-furanone
(rofecoxib) belongs to the group of NSAIDs known as COX-2 selective
inhibitors or coxibs (CycloOXygenase-2 InhiBitors). Being COX-2 selective
means that these drugs act preferentially on one form of the
cyclooxygenase (COX) enzyme, namely the COX-2, whereas earlier NSAIDs
inhibited both COX-1 and COX-2 with little if any selectivity. This
specificity allows rofecoxib and other COX-2 inhibitors to reduce
inflammation and pain while minimizing undesired gastrointestinal adverse
effects, e.g. peptic ulcers are common with non-selective NSAIDs such as
aspirin, naproxen, and ibuprofen.

[0009] Moreover it has been shown that there is an increased risk of
cardiovascular events associated with the use of rofecoxib, valdecoxib
and parecoxib and these compounds were withdrawn from the market.

[0010] Nevertheless, nonsteroidal anti-inflammatory drugs (NSAIDs) have
been successfully administered to treat pain and inflammation for many
years. The analgesic, antipyretic and anti-inflammatory properties of
aspirin and other NSAIDs were explained by the inhibition of prostanoid
synthesis by suppressing cyclooxygenase (COX) activity. This finding not
only explains the mechanism of action of NSAIDs, but also reveals a
useful pharmacological tool for evaluating the physiological role of
prostanoids by COX inhibition. NSAIDS are the most frequently used drugs
worldwide, based on the fact that they are administered to treat large
numbers of patients for many systemic pathological conditions, including
chronic polyarthritis, psoriatic arthritis, ankylosing spondylitis,
osteoarthritis, gout, inflammatory soft tissue rheumatism, lower back
pain, post-operative and post-traumatic inflammation, thrombophlebitis,
vasculitis, and certainly rheumatoid arthritis Dermatologic conditions
including erythema nodosum, nodular acne, prurigo nodularis, palmoplantar
pustulosis, and psoriasis that could lead to psoriatic arthritis, have
also been treated by NSAIDs. However, as noted above, long-term use of
NSAIDs is associated with gastrointestinal (GI) erosion and renal
failure.

[0011] Prostanoids (PGs) are widely distributed throughout the
gastrointestinal tract. Inhibition of PG synthesis is the principal
underlying mechanism for GI mucosal erosion. Endogenous prostaglandin
E2 (PGE2), derived from both COX-1 and COX-2, has been shown to
be involved in mucosal defense by decreasing gastric acid secretion,
which is mediated by prostaglandin E2 receptor (EP) type 3 (EP3) in
rats. PGE2 has a dual action on gastric acid secretion in rats, with
the inhibitory effect mediated by EP3 receptors and the stimulatory
effect through EP4 receptors. In mice, EP3 but not EP1
receptors are essential for acid-induced duodenal HCO3secretion and mucosal integrity. Endogenous PGI2, also has a
protective role in gastric mucosal integrity in mice. Unlike in rodents,
EP1 receptors are not found in any type of cells in the human
gastric mucosa.

[0012] In the kidney, prostanoids uphold the balance between
vasodilatation and vasoconstriction. They also regulate renin secretion,
tubular transport processes, and cell fate. PGI2 signaling is
essential for maintenance of renal homeostasis and renal function, which
is supported by the fact that COX-2 and PGI synthase (PGIS) knockout
mouse models display significant disturbances in the kidney. The
preferential association between COX-2 and PGIS in PGI2 biosynthesis
was evidenced by the fact that the renal phenotype of PGIS knockout mice
closely resembles the one shown in COX-2 knockout animals, while no major
abnormalities in COX-1 knockout mice were apparent. The same association
is also presented in humans. On the other hand, PGE2 regulates renal
hemodynamics and salt and water excretion via prostanoid receptors
EP1-4, where EP1 and EP3 act as constrictors, and EP2
and EP4 as dilators. The bidirectional capacity of PGE2 to
modulate vascular tone and epithelial transport allows PGE2 to serve
as a buffer to prevent physiological disturbances. Therefore, both
PGE2 and PGI2 are crucial prostanoids in regulating normal
kidney function. Their inhibition by NSAIDs results in sodium retention
and hypertension, which could cause acute renal failure.

[0013] Cyclooxygenases (COXs), COX-1 and COX-2, also known as prostanoid H
synthases (PGH synthase), are the key enzymes in the synthesis of
prostanoids from arachidonic acid released from membrane phospholipids.
The major difference between these two enzymes remains that COX-1 is
constitutively expressed as a "housekeeping enzyme" involved in
physiological functions in many cells, whereas COX-2 is usually expressed
inducibly and transiently. Because of the expression patterns of the two
isoforms of COX, it was assumed that COX-1-derived prostanoids were
involved in regulating physiological functions, whereas COX-2-derived
prostanoids played a major role in inflammation or tissue damage.
According to this hypothesis, the pharmacological effects of NSAIDs
depend on the inhibition of COX-2, whereas the toxic organ-specific
effects in GI tissue and kidney are linked to the inhibition of COX-1.
Therefore, as noted above, COX-2 selective inhibitors (COXIBs) were
developed as anti-inflammatory agents to minimize the side effects
associated with NSAIDs. However, in reality, COX-2 also plays a
physiological role in certain tissues and organs, and COX-1 may be
involved in inflammatory reactions. "Constitutive" expression has also
been observed for both isoforms of COX in the kidney, spinal cord, and
brain.

[0014] While there is no obvious advantage of COXIBs over nonselective
NSAIDS in terms of renal toxicity, significantly fewer GI complications
have been reported in patients treated with COXIBs. However, selective
inhibition of COX-2 causes an imbalance between COX-2 derived PGI2
and COX-1 derived TXA2, which results in serious cardiovascular
risk. TXA2 is a major prostanoid released from activated platelets
by COX-1 to stimulate platelet aggregation and vasoconstriction. To
counter the biological effects of TXA2, PGI2, a dominant
product of COX-2 under physiological conditions in vascular endothelial
cells, acts as a protective constraint on thrombogenesis, hypertension,
and atherogenesis. The imbalance in favor of TXA2 resulting from the
clinical use of COXIBs disrupts vascular homeostasis and, thus increases
vulnerability to thrombosis, atherosclerosis, and hypertension;
particularly in patients genetically susceptible to cardiovascular
disease. The withdrawal of the selective COX-2 inhibitors rofecoxib and
valdecoxib have emphasized the cardioprotective role of prostacyclin.

[0015] Although clearly demonstrated by the long and wide use of the
NSAIDs/COXIBs that prostanoids are critical inflammatory mediators, each
prostanoid, PGD2, PGF2α, PGI2, TXA2, and
particularly PGE2, might sometimes be anti-inflammatory. Most
importantly, the cardiovascular, renal, and gastrointestinal toxicities
associated with the NSAIDs/COXIBs must be addressed. It seems like that
EP3 receptors should be spared from blockade because of their roles
in GI protection, and IP receptors should be preserved because of their
importance in cardiovascular and renal homeostasis, while it might be
necessary to block TP receptors because of their cardiovascular liability
in clinical use of COXIBs.

[0016] Prostamide antagonist are known from published US Patent
Application Nos. 2008/0696240, 2005/0054699 and 2006/0106078. PGD2
antagonists are known from published US Patent Application No.
2004/0162333.

[0017] An object of the present invention is to provide compounds that
have FP, DP, EP1, EP4 and TP inhibitory activity, but lack EP2, EP3 and
IP activity and are useful as pharmaceuticals.

SUMMARY OF THE INVENTION

[0018] The present invention provides a method for treating inflammation
in a patient in need thereof comprising administering to said patient an
effective amount of a compound according to formula I

##STR00002##

wherein R is H or lower alkyl, R1 is hydrocarbyl or substituted
hydrocarbyl and the broken line represents a saturated or unsaturated
bond, i.e a double bond. Preferably R1 is an alkyl. More preferably R1 is
a n-alkyl or a cycloalkyl-n-alkyl, e.g. a cyclohexyl-n-alkyl, e.g.
n-octyl, n-nonyl or cyclohexyl-n-butyl radical.

[0019] The inventors of the present application have found that compounds
represented by the general formula (I) and prodrugs, isomers,
pharmaceutically acceptable salts thereof, i.e. acid and basic salts
thereof, have anti-inflammatory action and/are effective in reducing the
secretion of certain inflammatory cytokines and/or chemokines, making
them useful as pharmaceuticals. It has been shown that simultaneous
blockade of multiple potentially pro-inflammatory prostanoid receptors by
the compounds of this invention is either superior or at least as good as
rofecoxib (a COXIB) and diclofenac (a NSAID) in inhibiting the release of
multiple pro-inflammatory cytokines and chemokines The compound of
general Formula I blocks the following receptors: DP1, EP1, EP4, FP and
TP; and produces weak blockade of DP2.

[0020] The present invention provides a pharmaceutical composition
comprising the compound of formula I and a pharmaceutically-acceptable
carrier, in a solid or liquid dosage form, such as tablets, capsules,
powders, granules, suppositories, solution, suspension or emulsion.

[0021] Said pharmaceutical composition of the present invention may
further comprise one or more excipients, disintegrators, lubricants,
binders, preservatives, stabilizers and/or osmotic pressure regulating
agents.

[0023] Finally, the present invention provides certain novel compounds,
which are (E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[-
2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acids and lower alkyl
esters thereof. In the above formula, the novel compounds are represented
by those compounds wherein the alpha chain comprises a double bond.

[0024] Some of embodiments of the invention include but are not limited
to:

[0025] What is claimed is:

[0026] 1. A method for treating inflammation in a patient in need thereof
comprising administering to said patient an effective amount of a
compound according to formula

##STR00003##

wherein R is H or lower alkyl, R1 is hydrocarbyl or substituted
hydrocarbyl and the broken line represents a saturated or unsaturated
bond, a double bond, a wherein the formula includes prodrugs, isomers and
pharmaceutically acceptable salts thereof.

[0027] 2. The method of paragraph 1 wherein said compound is

##STR00004##

[0028] 3. The method of paragraph 2 wherein said method comprises
decreasing the secretion of ENA-78 in a patient.

[0029] 4. The method of paragraph 2 wherein said method comprises
decreasing the secretion of IL-8 in a patient.

[0030] 5. The method of paragraph 2 wherein said method comprises
decreasing the secretion of MCP-1 in a patient.

[0031] 6. The method of paragraph 2 wherein said method comprises
decreasing the secretion of PAI-1 in a patient.

[0032] 7. The method of paragraph 2 wherein said method comprises
decreasing the secretion of CD-40 in a patient.

[0033] 8. The method of paragraph 2 wherein said method comprises
decreasing the secretion of G-C--SF in a patient.

[0034] 9. The method of paragraph 2 wherein said method comprises
decreasing the secretion of GM-CSF in a patient.

[0035] 10. The method of paragraph 2 wherein said method comprises
decreasing the secretion of IL-1α in a patient.

[0036] 11. The method of paragraph 2 wherein said method comprises
decreasing the secretion of IL-18 in a patient.

[0037] 12. The method of paragraph 2 wherein said method comprises
decreasing the secretion of MDC in a patient.

[0038] 13. The method of paragraph 2 wherein said method comprises
decreasing the secretion of RANTES in a patient.

[0039] 14. The method of paragraph 2 wherein said compound is at least as
effective as COXIBs and NSAIDs in treating inflammation in a patient in
need of such treatment, without causing cardiovascular, renal and/or
gastro-intestinal side effects.

[0040] 15. A method for decreasing the secretion of the cytokine selected
from the group consisting of ENA-78, IL-8, MCP-1, PAI-1, TNFα,
CD-40, G-CSF, GM-CSF, IL-1α, IL-18, MDC and RANTES in a patient in
need thereof comprising administering to a patient an effective amount of
a compound according to formula I:

##STR00005##

wherein R is H or lower alkyl, R1 is hydrocarbyl or substituted
hydrocarbyl and the broken line represents a saturated or unsaturated
bond, such as a double bond.

[0041] 16. The method of paragraph 15 wherein said compound is:

##STR00006##

[0042] 17. A method according to paragraph 15 wherein said cytokine is
ENA-78 and said compound is administered for treating rheumatoid
arthritis.

[0043] 18. A method according to paragraph 15 wherein said cytokine is
IL-8 and said compound is administered for treating rheumatoid arthritis.

[0044] 19. A method according to paragraph 15 wherein said cytokine is
MCP-1 and said compound is administered for treating inflammatory
diseases characterized by monocytic infiltration, wherein such diseases
are selected from the group consisting of RA rheumatoid arthritus,
psoriasis, and atherosclerosis; atopic dermatitis, renal disease;
pleurisy; allergy and asthma; colitis; endometriosis; polymyositis and
dermatomyositis; uveitis; restenosis; brain inflammation and obesity;
diabetes and diabetes-induced atherosclerosis and MCP-1/CCR2-mediated
multiple inflammatory diseases.

[0051] 26. A method according to paragraph 15 wherein said cytokine is
RANTES and said compound is administered for treating rheumatoid
arthritis, inflammation, thrombosis and atherosclerosis, asthma,
including allergic lung inflammation, lung leukocyte infiltration,
bronchial hyper-responsiveness, and the recruitment of eosinophils in the
pathogenesis of asthma, and allergic rhinitis, multiple sclerosis, CNS
disorders, parasitic disease, cancer, autoimmune and heart diseases.

[0052] 27. A method according to paragraph 15 wherein said cytokine is
PAI-1 and said compound is administered for treating diseases relating to
the inhibition of plasminogen fibrinolysis, excessive fibrin
accumulation, and the sequential luminal obstruction, thrombus formation,
atherosclerotic lesions, vascular inflammation, and pathological
evolution of atherosclerotic plaques, deep vein thrombosis,
atherosclerosis, renal and pulmonary fibrosis, and cancer; as well as
metabolic syndrome with a combination of obesity, insulin resistance,
hypertension, hypertriglyceridemia, renal and cardiovascular disease.

[0053] 28. A method according to paragraph 15 wherein said cytokine is
TNFα and said compound is administered for treating diseases
resulting from the stimulation of the production of proinflammatory
cytokines/chemokines, collagenases, metalloproteinases, and other
inflammatory mediators; activation of endothelial cells and neutrophils;
promotion T- and B-cell growth, as well as stimulation of bone
resorption, the production of local and systemic proinflammatory
cytokines/chemokines and serum MMP-3, nitric oxide synthase activity,
VEGF release, and angiogenesis in inflamed joints.

wherein R is H or lower alkyl, R1 is hydrocarbyl or substituted
hydrocarbyl and the broken line represents a saturated or an unsaturated
bond, a double bond, and the compound includes prodrugs, isomers and
pharmaceutically acceptable salts thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0057] FIG. 1 shows a comparison of a representative compound of this
invention, i.e. the compound of Example 1, with diclofenac and rofecoxib
in modulating the secretion of ENA-78 from human macrophages stimulated
by LPS and TNFα (n=three donors, normalized by cell viability);

[0058] FIG. 2 shows a comparison of this representative compound of this
invention with diclofenac and rofecoxib in modulating the secretion of
IL-8 from human macrophages stimulated by LPS and TNFα (n=three
donors, normalized by cell viability);

[0059] FIG. 3 shows a comparison of this representative compound of this
invention with diclofenac and rofecoxib in modulating the secretion of
MCP-1 from human macrophages stimulated by LPS and TNFα (n=three
donors, normalized by cell viability);

[0060] FIG. 4 shows a comparison of this representative compound of this
invention with diclofenac and rofecoxib in modulating the secretion of
PAI-1 from human macrophages stimulated by LPS and TNFα (n=three
donors, normalized by cell viability);

[0061] FIG. 5 shows a comparison of this representative compound of this
invention with diclofenac and rofecoxib in modulating the secretion of
TNFα from human macrophages stimulated by LPS (n=three donors,
normalized by cell viability);

[0062] FIG. 6 shows the results a comparison of this representative
compound of formula I, diclofenac, and rofecoxib on CD-40 secretion from
human macrophages stimulated by LPS and TNFα (n=three donors,
normalized by cell viability);

[0063] FIG. 7 shows the results a comparison of this representative
compound of formula I, diclofenac, and rofecoxib on G-CSF secretion from
human macrophages stimulated by LPS and TNFα (n=three donors,
normalized by cell viability);

[0064] FIG. 8 shows the results a comparison of this representative
compound of formula I, diclofenac, and rofecoxib on GM-CSF secretion from
human macrophages stimulated by LPS and TNFα (n=three donors,
normalized by cell viability);

[0065] FIG. 9 shows the results a comparison of this representative
compound of formula I, diclofenac, and rofecoxib on IL-1α secretion
from human macrophages stimulated by LPS and TNFα (n=three donors,
normalized by cell viability);

[0066] FIG. 10 shows the results a comparison of this representative
compound of formula I, diclofenac, and rofecoxib on IL-18 secretion from
human macrophages stimulated by LPS and TNFα (n=three donors,
normalized by cell viability);

[0067] FIG. 11 shows the results a comparison of this representative
compound of formula I, diclofenac, and rofecoxib on MDC secretion from
human macrophages stimulated by LPS and TNFα (n=three donors,
normalized by cell viability);

[0068] FIG. 12 shows the results a comparison of this representative
compound of formula I, diclofenac, and rofecoxib on RANTES secretion from
human macrophages stimulated by LPS and TNFα (n=three donors,
normalized by cell viability);

[0069] FIG. 13 shows a synthetic scheme for the production of the
compounds of the present invention;

[0070] FIG. 14 is a summary of the results obtained in a comparison of a
representative compound of the compounds of the present invention and
various other PG antagonists in an in-vivo model of inflammation; and,

[0071] FIG. 15 is a summary of the results obtained in a comparison of a
representative compound of the compounds of the present invention and
various other PG antagonists in an in vivo model of neovascularization.

DETAILED DESCRIPTION OF THE INVENTION

[0072] Accordingly, the present invention relates to a method of treating
inflammation resulting from inflammatory diseases characterized by
monocytic infiltration caused by the secretion of cytokines and/or
chemokines by administration, to a patient in need of said treatment, of
a pharmaceutical composition comprising a compound represented by the
general formula:

##STR00009##

wherein R is H or lower alkyl, R1 is hydrocarbyl or substituted
hydrocarbyl and the broken line represents a saturated or unsaturated
bond, i.e. a double bond, in an amount effective to decrease the
secretion of cytokines and/or chemokines Preferably, R1 is an alkyl. More
preferably, R1 is a n-alkyl or a cycloalkyl-n-alkyl, e.g. a
cyclohexyl-n-alkyl, e.g. n-octyl, n-nonyl or cyclohexyl-n-butyl radical
and prodrugs, isomers and pharmaceutically acceptable salts thereof. Most
preferably, R1 is a cyclohexyl-n-butyl radical.

[0073] In the definition of the compound represented by the general
formula:

[0074] "Pharmaceutically acceptable salt" refers to those salts which
retain the biological effectiveness and properties of the free base and
which are obtained by reaction with inorganic acids such as hydrochloric
acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid,
methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,
salicylic acid and the like or retain the biological effectiveness and
properties of the free acid and which are obtained by reaction with
inorganic bases such as sodium and potassium hydroxides or carbonates,
etc.

[0075] "Prodrug" refers to compounds that decompose under in-vivo
conditions to yield the compound of formula I or a pharmaceutically
acceptable salt thereof.

[0076] "Alkyl" refers to a straight-chain, branched or cyclic saturated
aliphatic hydrocarbon. Preferably, the alkyl group has 1 to 12 carbons.
More preferably, it is an alkyl of from 4 to 10 carbons, most preferably
4 to 8 carbons. Typical alkyl groups include methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl and the like.
The alkyl group may be optionally substituted with one or more
substituents are selected from the group consisting of hydroxyl, cyano,
alkoxy, ═O, ═S, NO2, halogen, dimethyl amino, and SH.

[0077] "Cycloalkyl" refers to a cyclic saturated aliphatic hydrocarbon
group. Preferably, the cycloalkyl group has 3 to 12 carbons. More
preferably, it has from 4 to 7 carbons, most preferably 5 or 6 carbons.

[0078] The compounds of the invention have the basic profile of a safer
COXIB replacement with minimal cardiovascular liability and renal
toxicity, wherein prostanoid DP1, EP1, EP4, FP, and TP
receptors were blocked, DP2 receptors were partially blocked, while
leaving EP2 receptors open, and having no antagonizing activity at
IP receptors crucial for cardiovascular safety, and very little activity
at EP3 receptors important for GI tract and renal protection.
Therefore, said compounds provide a novel approach for anti-inflammatory
therapy. With this profile, it is hypothesized that the compounds
utilized in the method of this invention, e.g.
(E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo-
[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid would exhibit
anti-inflammatory effects in two dimensions: (a) blocking proinflammatory
PG effects via receptors DP1, EP1, EP4, FP, and TP; (b)
allowing PGE2 to exert anti-inflammatory effects via EP2
receptors. The unique pharmacological profile of
(E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo-
[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid also identifies
itself as a safer replacement for NSAIDs and COXIBs in two dimensions,
(a) by allowing IP receptors to exert their function of lowering blood
pressure and preventing platelet aggregation, and (b) by keeping EP3
receptors open for normal renal function and GI tract cytoprotection.
These features are advantageously different from just non-selectively
inhibiting all PG production from both COX enzymes like the NSAIDs, or
blocking all PG synthesized by COX-2 like the COXIBs.

[0079] The current finding that (E)-3-(2R-{3R-[4-(4
Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]hept-2-ylmethyl}-4-fluor-
o-phenyl)-acrylic acid or propionic acid and alkyl esters thereof are
effective in attenuating the production of TNF family cytokines (CD40 and
TNFα), and the classical interleukin-1 (IL-1) family cytokines
(IL-1α and IL-18) is especially important. These cytokines exert a
broad spectrum of biological and pathological effects. They play key
roles in inflammation and RA pathogenesis by stimulating the release of
multiple proinflammatory cytokines, including themselves, through the
NFκB signaling pathway. Certain antagonists/antibodies for IL-1 and
IL-18 are currently under preclinical investigation for RA treatment,
while the TNFα antibodies are already available for RA treatment.
Although alleviating the symptoms of RA in 50-65% of patients, a
TNFα antibody is very expensive to use compared to chemically
synthesized small molecules, inconvenient to administer usually requiring
injections, and has been linked to tuberculosis, lymphoma, and other
adverse effects. Unlike a TNFα antibody that totally eliminates all
circulating TNFα in the system,
(E)-3-(2R-{3R-[4-(4-Alkylyl-carbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]h-
ept-2-yl methyl}-4-fluoro-phenyl)-acrylic acid or propionic acid and alkyl
esters thereof only attenuates the production of TNFα by inhibiting
proinflammatory PG receptors. Therefore the adverse effects associated
with a TNFα antibody in elevating infectious and cancerous tendency
is less likely.

[0080] Finally, the current finding that
(E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]hept-
-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid or propionic acid or the alkyl
esters thereof inhibit PAI-1 production in human macrophages is important
in cardiovascular and metabolic pathological conditions. In addition to
PAI-1, other biomarkers inhibited by
(E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]hept-
-2-yl and methyl}-4-fluoro-phenyl)-acrylic acid or propionic acid and
alkyl esters thereof that are known to be closely related to inflammatory
conditions, have also been implicated in thrombosis and atherosclerosis.
Cells capable of forming atherosclerotic plaque include monocytes,
macrophages, endothelial cells, smooth muscle cells, platelets and T
cells. These cells express CD40 receptors, and release CD40 ligand
(CD40L) upon thrombin stimulation. MDC secreted by monocytes and
macrophages has been shown to be a strong and rapid activator of platelet
aggregation and adhesion. Proinflammatory elements IL-1, TNF, RANTES, and
MCP-1 are also involved in the cascade of events in the early and late
stages of atherosclerosis. Plasma MCP-1 levels have been linked to
cardiovascular disease risk factors in clinical studies. Platelet
activation leads to the release of MIP-1α, RANTES, ENA-78, and
IL-8, which attract leukocytes and further activate other platelets.
These evidences provide a direct linkage between homeostasis, infection,
and inflammation and the development of atherosclerosis.
(E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]hept-
-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid and propionic acid and alkyl
esters thereof is able to target multiple biomarkers of inflammation,
thrombosis, and atherothrombosis simultaneously, which may confer
pharmaceutical potential on
(E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]hept-
-2-yl methyl}-4-fluoro-phenyl)-acrylic acid and propionic acid and alkyl
esters thereof in treating atherosclerosis and atherothrombosis. As a
result, (E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.-
2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid and propionic acid and
alkyl esters thereof is unlikely to be associated with cardiovascular
liability as in the case of the COXIBs, conversely it may even have a
beneficial effect on cardiovascular function.

[0081] In summary, because of their ability to suppress the synthesis of
some key proinflammatory cytokines/chemokines CD40, ENA-78, G-CSF,
GM-CSF, IL-1α, IL-8, IL-18, MCP-1, MDC, RANTES, and TNFα, as
well as the adipocytokine PAI-1, the compounds of FORMULA 1 are not only
at least as effective as COXIBs and NSAIDs in RA treatment, but also are
a safer therapy in RA treatment. They are also be a potential therapy for
cardiovascular diseases.

[0082] The compounds of this invention treat or prevent inflammation at
least in part by the decreasing the amount of the secretion of certain
cytokines and/or chemokines that result from the exposure of the patient
to a stimulant.

[0083] In particular, the secretion of IL-8, ENA-7, MCP-1, PAI-1,
TNFα, CD-40, G-CSF, GM-CSF, IL-1α, IL-18, MDC, and RANTES is
reduced in those instances where said secretions are triggered by
lipopolysaccharides (LPS) and or TNFα.

[0084] The compounds of the present invention can be administered orally
or parenterally. The dosage is 3 to 150 mg/kg per day for an oral
administration and 1 to 50 mg/kg per day for a parenteral administration.

[0085] When the compound is administered as a pharmaceutical, it can be
prepared using usual formulation techniques and used in a dosage form of
solid or liquid, such as tablets, capsules, powders, granules,
suppositories, solution, suspension or emulsion.

[0086] Further, in this case, additive components which are customarily
used for pharmaceutical preparations such as excipients, disintegrators,
lubricants, binders, preservatives, stabilizers, osmotic pressure
regulating agents and so forth can be used.

[0088] The preparation of the compound of the present invention will be
described below in more details based on the Examples. The Examples
evaluate the effect of the compounds of this invention on the following
biological entities and are predictive of the effect of the present
compounds in treating diseases and conditions associated with said
biological entities.

[0089] Epithelial neutrophil-activating protein 78 (ENA-78, or CXCL5) and
Interleukin-8 (IL-8, or CXCL8): function as potent chemoattractants and
activators of neutrophils, ENA-78 and IL-8 are produced concomitantly in
response to stimulation with either IL-1 or TNFα. They not only
account for a significant proportion of the chemotactic activity for
neutrophils in rheumatoid arthritis (RA) synovial fluids, but also are
potent angiogenic factors in the RA synovium.

[0090] Monocyte chemoattractant protein-1 (MCP-1, or CCL-2): is not only
believed to play a role in inflammatory diseases characterized by
monocytic infiltration, such as RA rheumatoid arthritus, psoriasis, and
atherosclerosis, but is also implicated in other diseases, such as atopic
dermatitis, renal disease, pleurisy, allergy and asthma, colitis,
endometriosis, polymyositis and dermatomyositis, uveitis, restenosis,
brain inflammation and obesity. MCP-1 also controls leukocyte trafficking
in vascular cells involved in diabetes and diabetes-induced
atherosclerosis. MCP-1 antibodies are potential therapeutic agents for
treating MCP-1/CCR2-mediated multiple inflammatory diseases.

[0091] Plasminogen activator inhibitor 1 (PAI-1) is a potent inhibitor of
plasminogen fibrinolysis. In addition to inhibiting fibrinolysis, PAI-1
also plays an important role in regulating cell proliferation, adhesion,
migration, and apoptosis of vascular smooth muscle cells and endothelial
cells. TNFα elevates both local and plasma concentrations of PAI-1,
which results in excessive fibrin accumulation, and the sequential
luminal obstruction, thrombus formation, atherosclerotic lesions,
vascular inflammation, and pathological evolution of atherosclerotic
plaques. Increase levels of PAI-1 have been associated with pathological
conditions, including deep vein thrombosis, atherosclerosis, renal and
pulmonary fibrosis, and cancer; as well as metabolic syndrome with a
combination of obesity, insulin resistance, hypertension, and
hypertriglyceridemia, which will eventually develop into type 2 diabetes
and atherothrombosis. Judging from the fact that several currently
available renal protective drugs have PAI-1 inhibitory activity, PAI-1
also plays a key role in renal fibrosis. Therefore, abnormal PAI-1
expression may is a useful therapeutic target for renal and
cardiovascular diseases.

[0092] Tumor necrosis factor α (TNFα): mainly secreted by
macrophages and recognized for its importance in activating the cytokine
cascade. TNFα stimulates the production of proinflammatory
cytokines/chemokines, collagenases, metalloproteinases, and other
inflammatory mediators; activates endothelial cells and neutrophils;
promotes T- and B-cell growth, as well as stimulating bone resorption.
The TNFα antibody infliximab not only decreases the production of
local and systemic proinflammatory cytokines/chemokines, but also reduces
serum MMP-3 production, nitric oxide synthase activity, VEGF release, and
angiogenesis in inflamed joints.

[0093] CD40 signaling include regulation of antibody production by B
cells, induction of T cell proliferation, and activation of
antigen-presenting cells, such as dendritic cells (Rizvi et al., 2008).
In monocytes/macrophages, binding of the CD40L induces the
multimerization of CD40 receptors and activation of the TNF
receptor-associated factor (TRAF) pathway. Pro-inflammatory
cytokines/chemokines, such as IL-1α/β, IL-8, MIP-1α, and
TNFα, are released as the result of CD40-CD40L interaction. A
polymorphism in the CD40 locus is associated with the rate of joint
destruction in patients with RA (van der Linden et al., 2009), and a
strong association of the CD40 gene with RA has been confirmed in a large
UK case-control study (Orozco et al., 2009). CD40 is also present in
cells involved in atherosclerosis, including macrophages, endothelial
cells and vascular smooth muscle cells (Rizvi et al., 2008). The
interaction between CD40 and CD40L triggers platelet activation, which is
crucial for inflammation, thrombosis and atherosclerosis (Rizvi et al.,
2008). The soluble form of CD40, which is detected in the culture media
of human macrophages in the current study, is presumably produced by
shedding from membrane-bound CD40 (Schwabe et al., 1999). Soluble CD40
levels are observed to be elevated in the serum of patients with chronic
renal failure (Schwabe et al., 1999), chronic liver diseases
(Schmilovitz-Weiss et al., 2004), Alzheimer's disease (Mocali et al.,
2004), and systemic sclerosis--an autoimmune disease (Komura et al.,
2007).

[0094] Granulocyte colony-stimulating factor (G-CSF) is a major regulator
of neutrophil production and survival. G-CSF, along with
Granulocyte-macrophage colony-stimulating factor (GM-CSF), were the first
cytokines found in RA synovial fluid and synovium. The normally very low
serum levels of G-CSF rise dramatically during bacterial infection and
fall rapidly with resolution of infection. The elevation of serum levels
of G-CSF also correlate with disease activity of RA (Cornish et al.,
2009). G-CSF treatment can exacerbate underlying inflammatory diseases in
humans and mice, and G-CSF deficiency in collagen-induced arthritic mice
has profound protective effects. These findings suggest that G-CSF is an
important proinflammatory cytokine (Eyles et al., 2006).
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is produced by
a variety of cell types, including activated T cells and macrophages.
Production of GM-CSF can be stimulated by lipopolysaccharide (LPS), tumor
necrosis factor (TNF), and IL-1 (Cornish et al., 2009). While G-CSF is
largely neutrophil specific, GM-CSF stimulates the growth and
differentiation of neutrophils, macrophages, dendritic cells,
eosinophils, and erythrocytes [(Cornish et al., 2009). In addition to the
colony-stimulating effects on bone-marrow progenitor cells, a major role
for both G-CSF and GM-CSF is to enhance the production of mediators by
mature neutrophils and macrophages. Both G-CSF and GM-CSF are found in
the joints of RA patients. Administration of both cytokines exacerbates
RA, and antagonism of G-CSF or GM-CSF markedly reduces established
disease in mouse RA models. Biologic-based antagonists of both G-CSF and
GM-CSF are currently being developed and evaluated for RA therapy
(Cornish et al., 2009).

[0095] Interleukin-1α (IL-1α) and Interleukin-1β
(IL-1β) play an important role in immune regulation and inflammatory
processes by inducing nitric oxide synthase and matrix
metalloproteinases, as well as proinflammatory cytokines/chemokines
(Barksby et al., 2007) including TNFα, IL-8, ENA-78, MCP-1,
MIP-1α, and MIP-1β (St. Clair et al., 2004). Produced by
synovial tissue macrophages, activated T cells, fibroblasts and
chondrocytes, IL-1β is elevated in synovial fluids from RA patients
and results in the local effects of increased leukocyte infiltration and
MMP-mediated tissue turnover (Barksby et al., 2007). IL-1 cytokines are
key immune mediators responsible in inflammation and tissue destruction
in RA, and are detected after the early stages of RA (Barksby et al.,
2007). Clinical studies indicate that the recombinant human IL-1 receptor
antagonist Anakinra is a safe and well tolerated therapy for long-term
use in RA (Barksby et al., 2007).

[0097] Macrophage-derived chemokine (MDC) induces chemotaxis for
monocyte-derived dendritic cells, activated T cells and natural killer
(NK) cells (Ho et al., 2003). Highly expressed by the three major cell
types involved in allergic inflammation: eosinophils, basophils, and Th2
lymphocytes (Garcia et al., 2005), as well as highly expressed in atopic
dermatitis (Pivarcsi et al., 2005), MDC plays a role in inflammatory
diseases such as allergic asthma and atopic dermatitis (Ho et al., 2003).
Significantly enhanced in keratinocytes of patients with atopic
dermatitis, MDC could be a candidate therapeutic target for inflammatory
skin disease such as atopic dermatitis (Qi et al., 2009). MDC is also
implicated in disease activity of RA. After combination treatment with
the disease-modifying anti-rheumatic drugs leflunomide and methotrexate
in RA patients, plasma MCP-1 and MDC concentrations were significantly
lower, and so was the recruitment of inflammatory cells into the sites of
inflammation (Ho et al., 2003). Moreover, MDC also amplify platelet
activation and has been associated with the pathogenesis of
atherosclerotic disease including thrombosis (Gleissner et al., 2008).

[0098] Regulated on Activation, Normal T Cell Expressed and Secreted
(RANTES) is a chemoattractant for blood monocytes, memory T-helper cells
and eosinophils, and plays an active role in recruiting leukocytes into
inflammatory sites. It also stimulates the release of histamine from
basophils, activates eosinophils and causes hypodense eosinophils, which
is associated with diseases such as asthma and allergic rhinitis. RANTES
receptor CCR5 is also expressed on cells involved in atherosclerosis
(e.g. monocytes/macrophages, T lymphocytes, or Th1-type cells), and is
specialized in mediating RANTES-triggered atherosclerotic plaque
formation (Zernecke et al., 2008). Like MCP-1, stimulation with RANTES
enhances production of IL-6 and IL-8 in RA fibroblast-like synovial
cells; elevated MMP-3 production by chondrocytes, and inhibited
proteoglycan synthesis and enhanced proteoglycan release from the
chondrocytes (Iwamoto et al., 2008). Both MCP-1 and RANTES were found to
play an important role in allergic lung inflammation, lung leukocyte
infiltration, bronchial hyper-responsiveness, and the recruitment of
eosinophils in the pathogenesis of asthma (Conti et al., 2001). Similar
to MCP-1, RANTES also enhances the inflammatory response within the
nervous system, which plays an apparent role in the pathogenesis of
multiple sclerosis (Conti et al., 2001) Inhibitors for RANTES may provide
clinical benefits in treating inflammation, CNS disorders, parasitic
disease, cancer, autoimmune and heart diseases (Castellani et al., 2007).

[0099] Certain of the compounds utilized in the pharmaceutical
compositions and methods of treatment of the present invention are
prepared as shown in the following examples which examples are not
intended to be limiting but are preferred embodiments of the invention.

[0102] To a solution 2-bromo-4-fluorobenzaldehyde (15.2 g, 74.9 mmol) in
toluene (80 ml) was added (1R,2R)-(-)-pseudoephedrine (13.6 g, 82 mmol)
and the resulting mixture was refluxed removing water using a Dean-Stark
trap for 16 h. The reaction was halted and cooled down to room
temperature. The solution was washed with citric acid solution (1M, 100
ml), saturated sodium bicarbonate solution (50 ml), brine (50 ml) and
dried (MgSO4). Then, it was filtered and the solvent was evaporated
under vacuum to give title compound as yellow oil. (26.2, yield=97%).

[0106] At the same time, in a separate three neck 1 litre round bottle
equipped with A condenser, dropping funnel and under A nitrogen
atmosphere, 1,2-dibromoethane (7.95 ml, 92.2 mmol) was added slowly to a
stirred suspension of magnesium (2.15 g, 92.2 mmol) in anhydrous THF (30
mL) maintaining constant reflux. Once the fizzing had stopped, anhydrous
THF (100 ml) was added to suspend the white solid MgBr2 and the
suspension was cooled down to -60° C. To this cooled suspension,
the lithium salt solution prepared above was added by canula. The
resulting mixture was warmed up to -15° C. and stirred for 30
minutes. Then, it was cooled down to -60° C. and a solution of
norcantharidin (13.8 g, 82.2 mmol) in anhydrous THF (50 ml) was added
dropwise over 15 minutes and the resulting solution was stirred for 30
minutes. After this time the mixture was warmed up to -30° C. and
stirred for 2.5 h. Then the mixture was cooled down to -60° C. and
quenched with methanol (100 ml), followed by portion wise addition of
sodium borohydride (3.9 g, 101.9 mmol). The mixture wasallowed to warm up
to -25° C. and stirred for 1.5 h. A solution of hydrochloric acid
(2M, 150 ml) was carefully added, the mixture was warmed up to room
temperature and stirred for 14 h. The reaction mixture was concentrated
in vacuo diluted with water (100 ml) and extracted with EtOAc
(2×150 ml). The combined extracts were washed with brine (70 ml),
dried over MgSO4, filtered and the solvent was evaporated under
vacuum to give crude product as a green solid (19.5 g). The crude product
was purified by recrystallization from THF/isohexane (5:1) to yield the
title compound as a white solid (10.1 g, yield=50%).

[0115] To a solution of
3R-(5-Fluoro-2-formyl-benzyl)-7-oxa-bicyclo[2.2.1]heptane-2R-carboxylic
acid. (15 g, 54 mmol) in THF (500 mL) at rt and under nitrogen
atmosphere, (methoxycarbonylmethylene)triphenylphosphorane (27 g, 81
mmol) was added. The reaction mixture was stirred for 16 hours at rt
before concentrating in vacuo. The residue was dissolved in DCM
containing 10% of conc. NH4OH:EtOAc (1:9) (150 ml).
Triphenylphosphine oxide was removed from the crude product by filtering
the ammonium salt through 500 g of silica. The title compound was washed
out from silica using 10% AcOH in EtOAc and concentrated in vacuo to
yield an off-white solid (13.2 g, 73%).

[0119] To a solution of
3R-[5-Fluoro-2-(2-methoxycarbonyl-vinyl)-benzyl]-7-oxa-bicyclo[2.2.1]hept-
ane-2R-carboxylic acid. (13.2 g, 39.5 mmol),
2-amino-N-(4-cyclohexyl-butyl)-3-hydroxy-propionamide (10.5 g, 43.5 mmol
and NMM (6.8 ml, 59.3 mmol) in DCM (500 mL) with ice bath cooling, WSC
HCl (11.4 g, 59.3 mmol) was added. After 30 minutes the ice bath was
removed and the mixture was stirred for 16 hours at rt before
concentrating in vacuo. The residue was dissolved in EtOAc washed with
HCl (aq. 2M), sat. NaHCO3 and brine. The extract was dried over
MgSO4, filtered and the solvent was evaporated under vacuum to give
crude product as an off-white solid (12.5 g,).

[0132] The preparation of the compounds of this invention, as disclosed
above, wherein the cyclobutyl group is replaced by an octyl or nonyl
group are made by substitution of the appropriate reactant. The
structures of the compounds of the nonyl and octyl derivatives are given
in the SAR Table, as Examples 2 and 3, below.

[0133] The saturated compounds of Examples 4 through 6 are prepared as
follows:

[0138] To a solution of
(E)-3-[4-Fluoro-2-((S)-5-oxo-4,10-dioxa-tricyclo[5.2.1.0*2,6]dec-3-yl)-ph-
enyl]-acrylic acid methyl ester (18.1 mmol) in a mixture 2:1
methanol/tetrahydrofuran (75 ml), palladium hydroxide (0.61 g) was added.
The flask was evacuated and then connected to a balloon filled with
hydrogen. The reaction was stirred at room temperature for 2 h, and a
second portion of palladium hydroxide (0.61 g) was added. The flask was
evacuated and then connected to a balloon filled with hydrogen. After
another 2 h, the balloon was removed and Celite (1 g) was added to the
mixture, which was stirred for 10 minutes. The mixture was filtered
through a Celite pad and the pad was washed with methanol (25 mL). The
filtrate was evaporated to provide yellow oil. The oil was dissolved in
dichloromethane (50 mL) and dried over MgSO4.

[0139] Then, the solution was filtered and concentrated in vacuo and the
residue was dissolved in ethyl acetate (60 mL) and treated with Darco KB
activated carbon by heating at reflux for 2 minutes and then cooling.
Celite (1.2 g) was added and the mixture stirred for 10 minutes and then
filtered through a pad of Celite. The pad was washed with ethyl acetate
(25 mL). The filtrate was evaporated and the residue was crystallized
from hot ethyl acetate (11.5 mL) and heptane (23 mL). After cooling to
room temperature, additional heptane (30 mL) was added and the mixture
was left to stand at 4° C. overnight.

[0140] Then, I filtered the solid and washed with more heptane and dried
under vacuo overnight yielding the titled compound as a colourless solid.
(5.36 g, 88%)

[0144] After this time the solution was concentrated under vacuum and the
residue was dissolved in ethyl acetate (100 mL). Then, it was washed with
a 2M HCl solution (100 mL), saturated solution of NaHCO3 (100 mL)
and dried over MgSO4. Filtration and concentrated in vacuum yield
the titled compound as thick oil.

[0147] To a solution of
3-{4-Fluoro-2-[3-(2-hydroxy-1-nonylcarbamoyl-ethylcarbamoyl)-7-oxa-bicycl-
o[2.2.1]hept-2-ylmethyl]-phenyl}-propionic acid methyl ester (14.86 mmol)
in dichloromethane (200 ml), at -78° C. and under nitrogen
atmosphere, DAST (3.93 mL, 29.72 mmol) was added and the resulting
mixture was stirred at room temperature for 2.5 h. After this time,
potassium carbonate (4.11 g, 29.72 mmol) was added and the solution was
stirred for another hour. Then saturated solution of NaHCO3 (200 mL)
was added and the mixture was extracted with ethyl acetate (200 mL).
Then, it was washed with Brine (150 mL), and dried over MgSO4.
Filtration and concentrated in vacuo yield the crude compound as thick
oil.

[0148] The residue was purified by column chromatography in silica using a
solvent gradient starting from Ethyl acetate/iso-hexane 1:1 to Ethyl
acetate/methanol 9:1 to isolate the titled compound as thick oil (4.7 g,
60%)

[0152] To a suspension of Copper bromide (6.27 g, 28.08 mmol) in
dichloromethane (90 mL), under nitrogen atmosphere and in a water bath,
was added HMTA (3.94 g, 28.08 mmol) followed by DBU (4.17 mL, 28.08 mmol)
and the resulting mixture was stirred for 15 minutes. Then, a solution of
(3-{4-Fluoro-2-[3-(4-nonylcarbamoyl-4,5-dihydro-oxazol-2-yl)-7-oxa-bicycl-
o[2.2.1]hept-2-ylmethyl]-phenyl}-propionic acid methyl ester (3.72 g, 7.02
mmol) in dichloromethane (40 ml) was added and the resulting mixture was
stirred at room temperature for 16 h.

[0153] After this time, the solution was concentrated under vacuum and the
residue was partitioned between ethyl acetate (100 mL) and 1:1 sat.
solution of NH4Cl and NH3 (100 mL). Then, the organic layer was
separated and washed with Brine (100 mL), and dried over MgSO4.
Filtration and concentrated in vacuo yield the crude compound as thick
oil.

[0154] The residue was purified by column chromatography in silica using
Ethyl acetate/iso-hexane 5:1 to isolate the titled compound as a yellow
solid (2.2 g, 60%)

[0158] To a solution of
3-{4-Fluoro-2-[3-(4-nonylcarbamoyl-oxazol-2-yl)-7-oxa-bicyclo[2.2.1]hept--
2-ylmethyl]-phenyl}-propionic acid methyl ester 7 (1.39 g, 2.63 mmol) in
tetrahydrofuran (40 ml) was added a solution of Lithium hydroxide (0.441
g, 10.52 mmol) in water (10 mL) and the resulting mixture was stirred at
room temperature for 16 h. After this time, the solution was partitioned
between ethyl acetate (100 mL) and 2M HCl solution (50 mL). Then, the
organic layer was separated and washed with Brine (50 mL), and dried over
MgSO4. Filtration and concentrated in vacuo yield the titled
compound as slight yellow solid. (1.24 g, 92%)

[0160] The preparation of the compounds of this invention, as disclosed
above, wherein the nonyl group is replaced by an octyl or
cyclohexyl-n-butyl group are made by substitution of the appropriate
reactant. The structures of the octyl and cyclohexyl-n-butyl derivatives,
i.e. Examples 5 and 6, are given in the SAR Table, below.

[0161] In view of the above Examples and as shown in FIG. 13, the present
invention provides a method for the preparation of
(E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]hept-
-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid and propionic acid and lower
alkyl esters thereof, e.g.
(E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo-
[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid,

[0163] Preferably, this method comprises hydrolyzing a solution of
(E)-3R(2R{3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2-
.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid methyl ester in THF
and MeOH in the presence of NaOH, at room temperature, acidifying the
resulting solution with HCl, extracting the acidified solution with DCM,
drying the extract over MgSO4, filtering and concentrating the dried
extract in vacuo and recrystallizing said vacuum concentrated extract
from diethyl ether to yield
E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxabicyclo[2-
.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid as a white solid.

is prepared by reacting a solution of
3R-[5-Fluoro-2-(2-methoxycarbonyl-vinyl)-benzyl]-7-oxa-bicyclo[2.2.1]hept-
ane-2R-carboxylic acid with
2-amino-N-(4-cyclohexyl-butyl)-3-hydroxy-propionamide and NMM and said
reaction may take place in DCM at 0° C.

[0165] The (E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-o-
xa-bicyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid methyl
ester solution in DCM may be treated with HCl, concentrated under vacuum,
dissolved in EtOAc, washed with HCl, saturated NaHCO3 and brine,
dried over MgSO4, filtered and evaporated under vacuum to yield a
crude (E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-b-
icyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid methyl ester
as an off-white solid.

##STR00029##

[0166] In view of the above Examples and as shown in FIG. 13, the present
invention provides a method for the preparation of
(E)-3-(2R-{3R-[4-(4-Alkylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2.2.1]hept-
-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid and propionic acid and lower
alkyl esters thereof, e.g.
(E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo-
[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid,

[0168] Preferably, this method comprises hydrolyzing a solution of
(E)-3R(2R{3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bicyclo[2-
.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid methyl ester in THF
and MeOH in the presence of NaOH, at room temperature, acidifying the
resulting solution with HCl, extracting the acidified solution with DCM,
drying the extract over MgSO4, filtering and concentrating the dried
extract in vacuo and recrystallizing said vacuum concentrated extract
from diethyl ether to yield E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarb
amoyl)-oxazol-2-yl]-7-oxabicyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)--
acrylic acid as a white solid.

is prepared by reacting a solution of
3R-[5-Fluoro-2-(2-methoxycarbonyl-vinyl)-benzyl]-7-oxa-bicyclo[2.2.1]hept-
ane-2R-carboxylic acid with
2-amino-N-(4-cyclohexyl-butyl)-3-hydroxy-propionamide and NMM and said
reaction may take place in DCM at 0° C.

[0170] The (E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-o-
xa-bicyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid methyl
ester solution in DCM may be treated with HCl, concentrated under vacuum,
dissolved in EtOAc, washed with HCl, saturated NaHCO3 and brine,
dried over MgSO4, filtered and evaporated under vacuum to yield a
crude (E)-3-(2R-{3R-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-b-
icyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-acrylic acid methyl ester
as an off-white solid.

[0172] In order to measure the response of Gs and Gi coupled
prostanoid receptors as a Ca2+ signal, chimeric G protein cDNAs were
used. Stable cell lines over-expressing human prostanoid DP1,
EP1-4, FP, IP, and TP receptors were established as described in
U.S. Ser. No. 61/410,153.

[0173] As shown in the TABLE, the compounds of the Examples are selective
for the DP, EP1, EP4, FP and TP receptors and not the EP2 or IP receptors
and thus have the desired biological profile described above.

[0174] The compound of Example 1 was also evaluated for activity in an
in-vivo Rat Experimental Autoimmune Uveoretinitis (EAU) model and a model
of retinal neovascularization.

[0176] Rodent models of EAU induced by major uveitis autoantigens,
S-antigen (S-Ag) or A-Antigen-M18 peptide and interphotoreceptor retinoid
binding protein (IRBP) or R16 peptide are used to identify the efficacy
of various molecule and biologic therapeutics and evaluate the safety of
anti-inflammatory therapies.

[0177] It has been found that immunization with 50 μg of soluble
retinal protein peptide (M18) or 30 μg of photoreceptor proteins
peptide (R16) formulated with Freund's complete adjuvant mycobacterium
tuberculosis H3Ra by footpad injection in rat causes robust and
reproducible development of EAU. The ocular inflammatory response in the
EAU eyes will initiate in one-week post administration, reach its peak at
14-16 days and then subside at three to four weeks. This model was used
for testing the compound of EXAMPLE 1 for anti-inflammatory efficacy.

[0178] Prostaglandins (PGE, PGF, and PGD) have a variety of physiological
effects including the activation of the inflammatory response. When
tissues are damaged, white blood cells flood to the site; Prostaglandins
are produced via the cyclooxygenase pathway (COX) and activated by
binding to their membrane receptors to regulate tissue cell responses. It
has been shown that 1) the expression of EP2, EP4, and FP
receptors is excited in human ciliary epithelial and ciliary muscle
cells, 2) inhibition of prostaglandin receptor reduces ocular
inflammatory reaction; and 3) the prostaglandin receptor antagonists
reduce the PGs mediated ocular diseases or conditions, such as acute and
chronic uveitis.

[0179] The anti-inflammatory effect of PG antagonist of EXAMPLE 1 (as
compared to vehicle, alone, the compound of EXAMPLE 8, which blocks DP1,
EP4, FP and TP receptors; SC-51322, which blocks the EP1 receptor, only,
and the combination of EXAMPLE 8 and SC-51322) was measured. In this
test, EAU is induced by administering M18 peptide of S-Ag by ip injection
from day -1 to 13 to rats. This test demonstrates that if the multiple
prostaglandin receptors involved in the chronic ocular inflammatory
diseases are blocked inflammatory symptoms are not manifested. (See
http://www.iuphardb.org/DATABASE/LigandDisplayForward?ligandId=1924 for
the description of SC-51322.)

[0180] As shown in FIG. 14 (Anti-inflammation-Summary) the compound of
EXAMPLE 1 obtains a better clinical score than any of the comparators
except the combination of
3-(2-{(1S,2R,3S)-3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bi-
cyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-propionic acid and SC-51322
and a lower amount of protein and invasive cells in the aqueous humor
than all of the comparators. In addition, since it has been found that
the combination of
3-(2-{(1S,2R,3S)-3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bi-
cyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-propionic acid and SC-51322
inhibits ocular inflammation, it is clear that the compound of EXAMPLE 1,
which has the same biological profile as the combination, would also be
effective for treating ocular inflammation. Furthermore, as
representative of the other compounds utilized in the pharmaceutical
compositions and methods of treatment of this invention, said other
compounds will also be useful for treating and/or preventing ocular and
other inflammatory conditions.

Example 8

[0181] 3-(2-{(1S,2R,3S)-3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7--
oxa-bicyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-propionic acid is a PG
antagonist which is selective for DP1, EP4, FP and TP receptors.

Example 9

[0182] The compound of Example 1 was also evaluated for activity in an
in-vivo Rat laser-induced neovascularization (CNV) model.

[0184] Prostaglandin E2 (PGE2), once viewed as the prototypical
mediator of inflammation, is now regarded as a promoter of neoplastic
growth and of neovascularization. Several studies have delineated the
molecular mechanisms utilized by PGE2 to induce proliferation.
PGE2 upon binding to its membrane receptor, belonging to the
classical G protein-coupled receptor family, activates a signal cascade
that through a complex array of intermediate steps (c-Src, PKC, Pyk2),
leads to the extracellular release of peptide ligands stimulating growth
factor receptors and producing tumor growth. In parallel, PGE2
transactivates the EGF receptor (EGFR) via an intracellular
phosphorylation cascade involving the protooncogene c-Src, which
magnifies the EGF angiogenesis drive.

[0185] The angiostatic effect of the PG antagonist of EXAMPLE 1 (as
compared to vehicle, alone, the compound of EXAMPLE 8, which blocks DP1,
EP4, FP and TP receptors; SC-51322, which blocks the EP1 receptor, only,
and the combination of EXAMPLE 8 and SC-51322) was measured. In this
test, compounds were formulated in 70% polyethylene glycol (PEG). CNV is
induced by laser burn. This test demonstrates that if the multiple
prostaglandin receptors involved in the chronic ocular inflammatory
diseases are blocked, neovascularization, just like the inflammatory
responses in the above EAU model, is also not manifested.

[0186] As shown in FIG. 15 (Angiostatic Efficacy--Prostaglandin Receptor
Antagonists on Rat Laser CNV) the compound of EXAMPLE 1 obtains a better
clinical score than any of the comparators except the combination of
3-(2-{(1S,2R,3S)-3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bi-
cyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-propionic acid and SC-51322.
In addition, since it has been found that the combination of
3-(2-{(1S,2R,3S)-3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bi-
cyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-propionic acid and SC-51322
inhibits neovascularization, it is shown again that the compound of
EXAMPLE 1, which has the same biological profile as the combination of
3-(2-{(1S,2R,3S)-3-[4-(4-Cyclohexyl-butylcarbamoyl)-oxazol-2-yl]-7-oxa-bi-
cyclo[2.2.1]hept-2-ylmethyl}-4-fluoro-phenyl)-propionic acid and SC-51322,
would also be effective for treating neovascularization. Furthermore, as
representative of the other compounds utilized in the pharmaceutical
compositions and methods of treatment of this invention, said other
compounds will also be useful for treating and/or preventing
neovascularization.

[0187] The present invention is not to be limited in scope by the
exemplified embodiments, which are only intended as illustrations of
specific aspects of the invention. Various modifications of the
invention, in addition to those disclosed herein, will be apparent to
those skilled in the art by a careful reading of the specification,
including the claims, as originally filed. It is intended that all such
modifications will fall within the scope of the appended claims.